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Published ahead of print on November 7, 2003, doi:10.1165/rcmb.2003-0260OC

Am. J. Respir. Cell Mol. Biol., Volume 30, Number 5, May 2004, 627-634

A more recent version of this article appeared on May 1, 2004
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Submitted on July 10, 2003
Revised on November 7, 2003

P.aeruginosa flagella activate airway epithelial cells through asialoGM1 and TLR2 as well as TLR5

Robert Adamo1, Sach Sokol1, Grace Soong1, Marisa Gomez1, and Alice Prince1*

1 Pediatrics and Pharmacology, Columbia University, New York, NY, United States

* To whom correspondence should be addressed. E-mail: asp7{at}columbia.edu.

The distribution of specific toll like receptors and components of the signaling pathways activated by P.aeruginosa flagella were studied in airway epithelial cells. Initially flagella bound to the apical surface of polarized epithelial cells where they prominently co-localized with asialoGM1. By 4 hours of exposure to flagella, TLR5 expression was induced, mobilized to the apical surface of the cells and co-localized with superficial flagella. IL-8 expression in airway cells was activated by flagella through induction of Ca2+ fluxes, Src, Ras and ERK1/2 MAPK and NF-{kappa}B activation, a pathway previously associated with asialoGM1-mediated stimuli. There was evidence for participation of asialoGM1 and TLR2 as well as TLR5 in the response to flagella and increased asialoGM1 correlated directly with increased signaling. TLR2 DN or TLR5 DN mutations inhibited IL-8 induction by 78% and 35% respectively (P<0.001 for each). The participation of TLR2 as well as TLR5 was confirmed in CHO cells transfected with either human TLR2 or TLR5 in which flagella activated an NF-{kappa}B-luciferase reporter to the same extent. Flagella signaling in airway cells can be initiated by interactions with asialoGM1 and TLR2 as well as by activation of TLR5. The availability of exposed receptors on the apical surface of polarized airway epithelial cells is a major factor in the activation of signaling pathways by flagella.




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